Molecular methodologies for improved genetic diversity assessment in cucumber and melon

J. E. Staub, M. D. Robbins, S. Chung, A. I. López-Sesé

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

3 Scopus citations

Abstract

Genetic markers have been used successfully in germplasm management to catalogue accession relationships and develop strategies for their management. Molecular marker technology has been applied to the assessment of genetic diversity of Cucumis species. The interpretation of relationships among accessions is made more meaningful if marker-based diversity assessments are standardized by uniform marker arrays (SMA) and historically based reference accessions (RA). In this study, previously mapped random amplified polymorphic DNA (RAPD) markers were converted to more stable sequence characterized amplified region (SCARs) markers and tested in multiplexing reactions (duplex and triplex) to increase assessment efficiency. Efficiency of PCR analysis can be increased by the use of liquid dispensing machines (LDM) that operate on a 96-well format. The efficiency of one such machine was compared to manual dispensing in preparation of PCR. The effectiveness (based on band intensity) of LDM and manual PCR preparation was equivalent. When compared to manual preparation, the efficiency of PCR preparation is increased using the LDM if more than four plates are run per day. Although a single SCAR marker can function well when used alone or in combination with another SCAR marker, the complementary use of markers in multiplex reactions decreases as the number of markers used in the polymerase chain reaction (PCR) increases. The kinetics of the PCR and its optimization play a major role in the success of multiplexing.

Original languageEnglish
Title of host publicationXXVI International Horticultural Congress
Subtitle of host publicationAdvances in Vegetable Breeding
PublisherInternational Society for Horticultural Science
Pages41-47
Number of pages7
ISBN (Print)9789066056671
DOIs
StatePublished - 31 May 2004

Publication series

NameActa Horticulturae
Volume637
ISSN (Print)0567-7572

Keywords

  • Multiplexing reactions
  • Reference array
  • Standard marker array

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